TW201206493A - Elastase inhibitor - Google Patents

Abstract

The present invention relates to an elastase inhibitor. The elastase inhibitor contains rubus extracts and hydroxyproline as active ingredients. Yeast extracts can be optionally further included in the ingredients. The elastase inhibitor can be utilized for medical treatment and skin care.

Description

201206493 VI. Description of the Invention: [Technical Field] The present invention relates to an elastase inhibitor comprising as an active ingredient a raspberry extract and hydroxyproline as a component of a plant. [Prior Art] 〇 〇 Previously, considering the need for anti-aging agents, J·sheng, but the mechanism or definition related to aging is not clear. Therefore, in general, the moistness of the skin is measured by moisturizing state. Or the determination of skin elasticity, or visually observing the color of the skin. However, in recent years, research related to aging has progressed. As a cause of skin aging, macro growth is an important factor, and drying, oxidation, and the influence of sunlight (ultraviolet rays) can also be cited as direct related to skin aging. factor. As a specific expression of skin aging, it is known that the collagen or elastin in the skin dermis is reduced, the mucopolysaccharide represented by hyaluronic acid is reduced, and the damage of cells caused by ultraviolet rays is caused. It is generally believed that elastin will crosslink with each other to contribute to the elasticity of the tissue, but due to ultraviolet exposure or age, elastase, which is an elastin-destroying enzyme, will be excessively expressed, resulting in modification and destruction of elastin. It is associated with a decrease in the elasticity of the skin. Because it inhibits the function of elastase, it prevents the modification of elastin that imparts elasticity or tension to the skin. Destruction is important to prevent skin aging. In the case of directly coating a cosmetic or the like on the skin, it is preferred to use a natural ingredient as such a natural elastase inhibitor, for example, known as the extract of the genus Linden (Indian linden) (Patent Literature) Saxifrage extract (Patent Document 2), Rubiaceae plant extract (specially 150170.doc 201206493 Literary Document 3), aunt Yu Xinshi, the death of the Rhododendron simsii ( Special & literature 4) etc. and reveals: skin external use containing the extract 1; wrinkles, wrinkles and tension of the skin. Relaxation shows improvement effect 0 Another v 'elastase inhibitor except skin external preparation It is also known to have usefulness as a therapeutic agent for diseases, such as reports: joint system diseases such as chronic rheumatoid $ human k-shaped joint disease, systemic inflammatory reaction

Symptoms, arteriosclerosis, I ~, axillary pulmonary mussel injury, acute respiratory distress syndrome, etc. are more effective. / Body and t 'The following pharmaceuticals are known: ulinastatin for acute vomiting or acute circulatory failure (hemorrhagic shock), and the like. An effective selective lung sputum associated with acute inflammatory response syndrome in the stagnation of the stagnation of spleen) Siveles sodium (3) (8)

Sodlum). For example, &' elastase inhibitor can be used as a therapeutic agent for inflammatory diseases and the like. However, in consideration of safety and the like, it is purely a product which is a component derived from a natural component other than a chemical composition. [Patent Document 1] [Patent Document 1] Japanese Patent Laid-Open Publication No. Hei. No. Hei. No. Hei. No. Hei. No. 11-246386 (Patent Document 3) Japanese Patent Laid-Open No. 824! [Patent Document 4] Japanese Patent Laid-Open Publication No. Hei. No. 4 (a) The object of the present invention is to provide an elastase inhibitor which can inhibit elastase by 150170.doc, 201206493: external: Thereby, the skin is given elasticity or tension and is also effective as a therapeutic agent for the disease. The inventors of the present invention have investigated the activity of the elastase-inhibiting activity of various plant extracts, and as a result, it has been found that the solvent extract of the squid (Rubusidacus L) has elastase inhibitory activity, and it is found that the inhibitory activity is obtained by adding the citrate. The invention has been completed by exponentially increasing. Therefore, the present invention provides the following invention. ❹ (1) An elastic egg (tetra) inhibitor comprising a fisherman extract and a transbasic acid as an active ingredient. (7) The elastase inhibitor of (1) wherein the above-mentioned transbasic acid is L-hydroxyproline. (3) The elastase inhibitor according to (1) or (2), which further comprises a yeast extract. (4) The elastase inhibitor according to (3), wherein the yeast extract is a yeast which is cultured in a nutrient medium containing glycosaminoglycan and treated by ultraviolet irradiation, hydrogen peroxide treatment or the like. And prepared. (7) A skin external application comprising the elastase inhibitor according to any one of (1) to (4). (6) The skin external preparation according to (5), which is a cosmetic. [Embodiment] Hereinafter, embodiments of the present invention will be described. The core hook (Raspberry, European Rubus) (Rubus idacus l) is a Rosaceae, Rubus plant. The deciduous shrubs distributed in the wilderness from Europe to West Asia and the east coast of the Americas have a flowering period of about 6 months. The fruit is ^ 〇 150170.doc 201206493 2 right mature 'below red and become purple with black. It is commonly known as raspberry, color, fragrance - representative fruit of the era, even now ^ and the result of rare fruit processing Sauce or syrup, etc. As a drug such as citric acid, such as sweet w έ ^ Knife 3 malic acid, citric acid and other sweet astringent substances, with estrogen-like activity such as Huang Gang 'to vitamin c (ascorbic acid), vitamin ρ (orange peel, Musk buds are various vitamins or saccharides of 2, and therefore, in addition to the effects of dermatitis and thin sputum, anti-inflammatory agents such as throat and diarrhea are also known. Examples of the raspberry extract used in the elastic egg self-enzyme of the present invention include a solvent extract, a diluent of the solvent extract, and a dried product obtained by drying the solvent extract, and drying the dried extract. A solution obtained by dissolving the substance in a solvent or the like. Further, the crude purified product, the purified product and the like are also included. The extraction solvent used in the present invention may be any solvent which is usually used for extraction, and may be an alcohol such as water, decyl alcohol, ethanol or hydrazine, 3-butanediol, or an alcohol such as acetone or ethyl acetate. The solvents are used singly or in combination, and particularly preferred are alcohols, aqueous alcohols, particularly preferably ethanol, hydrazine, 3-butanediol, aqueous ethanol or aqueous 1,3-butanediol. Further, the above solvent is preferably used at a temperature from room temperature to the boiling point of the solvent. As the site of the above plant body, it is preferred that the fruit is used, and an extract of other parts may be used. The raspberry extract used in the present invention has excellent elastic protease inhibitory activity against human skin, and thus the skin external application of the plant extract 150170.doc 201206493 can prevent skin aging or maintain a healthy skin condition. In the case where the elastase inhibitor of the present invention is formulated in an external preparation, the amount of the raspberry extract is 0.000001 to 1% by mass in the total amount of the external preparation, preferably 〇〇. 〇〇〇1 to 〇丨% by mass, more preferably 0.0001 to 0·01% by mass, most preferably about 〇001% by mass. If it is less than 0 '000001% by mass, the effect described in the present invention cannot be sufficiently exerted, and if it exceeds 1% by mass, it is difficult to form a preparation, which is not preferable. The elastase inhibitor of the present invention may also be a suspension containing only the raspberry extract, preferably containing hydroxyproline, more preferably a yeast extract. In particular, elastase inhibitory activity can be multiplied by the formulation of hydroxyproline. The amino acid, particularly L-hydroxyproline, is a collagen-specific amino acid. The amino acid sequence of collagen is -G1y_X_Y_, X, Y are any amino acids, and at least one contains L-valine or L-hydroxyproline. It has the effects of promoting the production of collagen in fibroblasts, proliferation of epidermal cells, moisturizing effects, and the like to prevent skin aging. In the case where the elastase inhibitor of the present invention is formulated in an external preparation, the amount of the hydroxyproline is adjusted to 0.00001 to 10% by mass, preferably 0.0001, based on the total amount of the external preparation. ~1 quality. More preferably, it is 0.001 to 0.1% by mass, and most preferably about 0.025% by mass. If it is less than 0.00001% by mass, the effects described in the present invention are not sufficiently exerted, and if it exceeds 10% by mass, it is difficult to form a preparation, which is not preferable. In the present invention, it is preferred that the yeast extract is used in a yeast culture medium which is cultured in a nutrient medium containing glucosamine polyfluorene and subjected to ultraviolet irradiation of 'hydrogen peroxide treatment or 150170.doc 201206493 or both. The presence of a stress such as chlorine peroxide is known to be due to the fact that the yeast is cultured in the lower part of the yeast, and the nutrient protein glands are added to the stress group. And the improvement of the stress = should be. The method of the method is described in, for example, U.S. Patent No. 6,461,857. For example, yeast extract can be prepared by the following method: Saccharomyces cerevisiae as a kind of baker's yeast (Saeeh_yees e(10)is (4) is cultured in a nutrient medium supplemented with glycosaminoglycan which is not derived from an animal, and a sublethal dose such as a mass of about 2 mass of the total mass of the culture is added to the culture. Further, the sub-lethal irradiation amount of ultraviolet rays (for example, UVA/UVB irradiation is set to an intensity of 31.5 mJ/cm 2 ) is applied arbitrarily, thereby stress is applied, and the resulting culture is obtained by self-decomposition or acid hydrolysis. After the substance is dissolved, it is dried, extracted with water, filtered, etc. Further, the yeast extract prepared by the yeast which is cultured in a medium containing not the glycosaminoglycan from the animal and treated with ultraviolet rays and hydrogen peroxide is used. Arch Personal Care Products LP is sold under the trade name "Biodyne EMpp (trademark)". When the elastase inhibitor of the present invention is formulated in an external preparation, the amount of the yeast extract is adjusted to the total amount of the external preparation. The dry matter is 0.000001 to 1% by mass, preferably 0.00001 to 0% by mass, more preferably 0.0001 to 〇.〇1% by mass, most preferably 0.0005 to 005% by mass. When 0.000001% by mass, the effect described in the present invention is not sufficiently exhibited. If it exceeds 1% by mass, it is difficult to form a preparation, and therefore it is not preferable. 0 150170.doc 201206493 In the case where the elastase inhibitor of the present invention is used as an external preparation It can be used with ingredients such as whitening agents, moisturizers, antioxidants, oily ingredients, and ultraviolet light absorption agents commonly used in cosmetics or pharmaceuticals. , Surfactants, thickeners, alcohols, powder components, a colored material, aqueous components, water, various skin nutrients and the like to be appropriately formulated in combination.

In addition, metal sulphate, tannin, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid and other metal blocking agents 'caffeine, tannin, etc. Verapamil, tranexamic acid and its derivatives, licorice extract, licorice, hot water extract of Pyracantha fortune, various Chinese herbal medicines, tocopherol acetate, glycyrrhizic acid and its derivatives or its salts 'Vitamin c, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, citric acid and other whitening agents, glucose, fructose, mannose, sucrose, trehalose and other sugars. The external preparation containing the elastase inhibitor of the present invention can be widely applied to cosmetics, quasi-drugs and the like which are applied to the epidermis of the skin, and is particularly preferably applied to cosmetics, and the dosage form can also be an aqueous solution or a solubilizing system. A wide range of dosage forms such as an emulsified umbrella blade, an oil system, a gel system, an ointment system, an aerosol system, and a water-oil two-layer water-oil-powder three-layer system. In other words, in the case of the above-mentioned various types (4), it can be widely used in the form of facial cleansers, lotions, lotions, creams, gels, serums (cosmetic liquids), masks, etc. X, if it is a makeup Cosmetics can be widely used in foundations such as foundations for showers, etc. Further, if it is a quasi-drug, it can be widely used in various ointments and the like, and the elastase inhibitor of the present invention is coated. The form of the external preparation may not be used: 150170.doc 201206493 is determined by the dosage form and form. Further, the elastase inhibitor of the present invention may be used, or acute lung injury..." ^ m 器m田和箸迫钱群(4) and other therapeutic agents for diseases such as stagnation agents. In the middle of the process, in order to use the elastase inhibitor preparation 2 of the present invention, for example, powders, granules, and lyophiles are produced by a usual method. Isotonic solution, etc., in the case of preparing a solid preparation for oral administration, after adding a binder, a wet full agent, a disintegrant, a surfactant, a dispersant, a flavoring agent, a deodorant, etc., as needed According to the usual gas per tablet, coated lozenge, granules, capsules, etc. Examples of the excipients used include lactose, glucose, sorbus/n temple powder, mannitol, etc., and binders, for example, Examples thereof include polyvinyl alcohol, polyvinyl ether, ethyl cellulose, gum arabic, gelatin, propyl cellulose, and polyethylene (four). Examples of the disintegrant include acid dance, citric acid, dextrin, and starch. The gelatin powder, etc., may be exemplified by a carbonated mother, a citric acid, a talc, a polyethylene glycol, etc., and may be exemplified by cocoa powder, mint aromatic acid, peppermint oil, etc. The granules need to be appropriately coated with sugar coating, gelatin coating and other knives: when preparing the injection, add pH adjustment as needed 4 buffer, surfactant, dissolution aid, solvent, stabilizer, Baoqi The preparation of subcutaneous, intramuscular, and intravenous injections according to the conventional method. The present invention provides, in a further aspect, the use of the elastase inhibitor of the present invention as a skin external preparation, preferably a cosmetic. The agent can be used in the following manner, that is, in order to suppress/pre-150170.doc -10·201206493, the elasticity of the skin accompanied by the modification and destruction of elastin is reduced, and the application is to suppress/prevent the above skin. In the subject of reduced elasticity, such as the skin thereof, the modification and destruction of the above elastin is caused by excessive expression of elastase caused by, for example, ultraviolet exposure or age increase. Further, the present invention provides the elastase inhibitor of the present invention. Uses for the treatment and/or prevention of elastase-related diseases such as chronic rheumatoid arthritis, articular diseases such as osteoarthritis, systemic inflammatory response syndrome, arteriosclerosis, acute lung injury, acute respiratory distress syndrome The elastase inhibitor of the present invention can be applied to a subject who needs to suppress/prevent the above-mentioned skin elasticity reduction by oral or parenteral administration. [Examples] Hereinafter, the present invention will be specifically described by way of examples, but the present invention is not limited to the following examples. As a reaction buffer, 0.1 M HEPES (4-(2-hydroxy ethyl) -1 -piperazine ethane sulfonic acid, 4-ethyl π bottom p well) and 〇 5 μ (pH 7.5) were used. Dissolving dms〇 (Dimethyl Sulphoxide, dimethyl) in an amount of 80 mM as an elastase matrix of oxydopyl-(Ala)2-Pro-Val-MCA (PEPTIDE INSTITUTE Cat No. 3153·ν) In Aachen), use the reaction buffer to dilute to 16 mM. Elastase from human leukocytes (ELASTIN PRODUCT CO., INC. Cat No. CK828) was diluted to 5 pg/mL using anti-application buffer. A 25 μL of 0.16 mM elastase matrix was dispensed into each of the 96-well plates, and 50 pL of each agent was added. Then, 5 pg/mi of elastin was added to ice 150170.doc -11 - 201206493 Enzyme 25 pL and cultured at 37 C for 60 minutes, and 460 nm fluorescence was measured at 369 nm. Using the buckwheat extract BG of Maruzen Pharmaceutical Co., Ltd. as the raspberry extract, using L-hydroxyproline as the hydroxyproline in Concord Industrial Co., Ltd., and using Biodyne of Arch Pers〇nal CareL p EMPP (trademark) as a yeast extract. The buckwheat extract BG is a 1,3 - butanediol (5 % aqueous solution) extract of the fruit of the fish. The results are shown in Figure i. Significant concentration-dependent elastase inhibition was confirmed by using the squid extract alone. On the other hand, the use of hydroxyproline did not exhibit an elastase inhibitory effect. Interestingly, it can be seen that although the block IW m4 male..., & glutamic acid did not show elastase inhibitory effect, but it can be significantly enhanced with the use of scorpion benzene smashing / i Η 〇 卒 卒 卒 卒The elastase of the raspberry extract inhibits the ~, pressing effect. Therefore, it can be seen that the elastase inhibiting effect of the cardiac hooks can be multiplied by the combination of gibberyl acid. Figure 2 shows that Μ!- onft... 单独 苟 — — — ( ( ( 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0.001 The amount of ° /.) and the fermented material (in terms of your 〇t is 0.001% by mass), the production of eight, 〖月形日卞, and the use of the open v% (in dry matter Ye Maqi, 〇〇1% by mass '〇〇.〇〇1% by mass of the elastic egg J. As the material as as the extraction of the raspberry extract - ... the effect of each agent is compared with the use of each of the early protease inhibitors. (4) The heart will be more prominently improved by the formula 1 cream (prescription) 150170.doc 201206493 (1) Stearic acid 3.0% by mass 5.0 18.0 3.0 10.0 0.01 0.001 0.2 0.01 Appropriate amount of the remaining (2) Stearyl alcohol (3) Isopropyl myristate (4) Glycerol monostearate (5) Propylene glycol (6) L-hydroxyproline (7) Raspberry extract (as dry) (8) Caustic potash

(9) Sodium bisulfite (10) Preservatives (11) Perfume (12) Ion exchange water (preparation method)

To the ion-exchanged water, propylene glycol, L i-surface valine acid, raspberry extract, caustic potash, and dissolved were added, and the crucible was added and maintained at 7 〇 C (aqueous phase). The pots, ingredients were mixed, heated to melt and held at 70. [] (oil phase). The oil phase is added to the water phase, and after the addition is completed, the temperature is maintained for a while: reaction. Thereafter, it was uniformly emulsified by a homomixer, and the surface was cooled to 30 ° C while being thoroughly stirred. Prescription Example 2 Cream (prescription) 2.0% by mass 7.0 3.0 (1) Stearic acid (2) Stearyl alcohol (3) Hydrogenated lanolin 150170.doc -13- 201206493 4.0 6.0 3.0 2.0 6.0 0.02 0.005 0.001 0.03 0.3 Appropriate amount (4) squalane (5) 2-octyldodecanol (6) polyoxyethylene (25 mol) green 4 murine alcohol linkage (7) glycerol monostearic acid vinegar (8) propylene glycol (9) L - Hydroxyproline (10) Elastase inhibitor of the elemase (as dry) Yeast extract (as dry) (11) Sodium bisulfite (12) Hydroxybenzoic acid (13) ) Perfume (14) Ion-exchanged water (preparation method) Propylene glycol is added to ion-exchanged water, heated and kept in a thief (aqueous phase). The other ingredients were mixed and heated to 7 〇t (oil phase). The oil phase was added to the aqueous phase and pre-emulsified, and after homogenization, the mixture was thoroughly stirred and cooled to 3 Torr. Formulation Example 3 Emulsion (Prescription) 2·5质量% 1.5 5.0 10.0 (1) Stearic acid (2) Cetyl alcohol (3) Vaseline (4) Liquid paraffin 150170.doc • 14 - 201206493 2.0 3.0 1.0 0.05 0.003 0.005 0.01 0.3 Appropriate amount of the remaining (5) polyoxyethylene (l〇 mol) monooleate (6) polyethylene glycol 15 〇〇 (7) triethanolamine (8) carboxyvinyl polymer (9) the elastase inhibitor L - Hydroxyproline sulphate extract (as dry) (10) Sodium bisulfite 11 (11) p-Hydroxybenzoate ethyl ester (12) Perfume (13) Ion exchange water (preparation method) in a small amount of ion exchange Dissolve the Wei ethylene polymer (A phase) in water. Polyethylene glycol 15 hydrazine and triethanolamine were added to the remaining ion-exchanged water, dissolved by heating and kept at 70 ° C (aqueous phase). The other ingredients were mixed, heated and melted and kept at 7 ° C (oil phase). The oil phase was added to the aqueous phase and pre-emulsified, and the phase A was added and uniformly emulsified by a homomixer, and after emulsification, the mixture was sufficiently stirred to be cooled to 30 °C. Formulation Example 4 Emulsion (Prescription) (1) Microcrystalline wax 1.0% by mass (2) Beeswax 2.0 (3) Lanolin 20.0 (4) Liquid sarcophagus 1〇〇150170.doc -15- 201206493 5.0 (5) Squalane (6) Sorbitol trioleate 4.0 (7) Polyoxyethylene (20 mol) Sorbitol 1.0 Anhydride monooleate (8) Propylene glycol 7.0 (9) Elastase inhibitor L-hydroxyguanamine 〇.〇1 Raspberry extract (as dry) 0.001 Yeast extract (as dry) 0.005 U〇) Syzygium leaf extract 〇.05 (11) Sodium bisulfite 〇.〇1 (12) Ethyl p-hydroxybenzoate 3.3 (13) Proper amount of perfume (14) Ion exchange water remaining (preparation method) Propylene glycol was added to ion-exchanged water, and heated and maintained at 70 C (aqueous phase). Mix the other ingredients 'heated and maintained at 7 〇 (oil phase). The oil phase was stirred on one side, and the aqueous phase was slowly added thereto, and emulsified uniformly by a homomixer. After emulsification, stir it to 3 (rc.) Prescription Example 5 Beauty Solution (Prescription) (A phase) 10.0% by mass 1.0 (1) Ethanol (95%) (2) Polyoxyethylene (20 mol) octyl ten Dialkyl alcohol 150170.doc -16· 201206493 (3) Panthenium ether 0.1 (4) Elastase inhibitor L-hydroxyproline 0.001 Raspberry extract (as dry) 0.001 (5) p-Hydroxybenzoic acid Methyl ester 0.15 (B phase) (6) Potassium hydroxide 0.1 (C phase)

(7) Glycerin 5.0 (8) Dipropylene glycol 10.0 (9) Sodium bisulfite 0.03 (10) Carboxyrene polymer 0.2 (11) Purified water remaining (preparation method) A phase 'C phase is uniformly dissolved, respectively, to phase C Add phase A and dissolve. Then add phase B and then fill it. Prescription Example 6 Lotion (Prescription) (1) Ethanol 5.0% by mass (2) Glycerol 0.5 (3) Dipropylene glycol 2.0 (4) 1,3-Butanediol 5.5 (5) Citric acid 0.02 (6) Sodium citrate 0.08 150170.doc -17- 201206493 (7) Sodium hexametaphosphate 0.03 0.1 0.003 0.003 0.1 0.001 Residual (8) Hydroxypropyl-β-cyclodextrin (9) Elastase inhibitor L-hydroxyproline dewberry extraction Liquid (as dry matter) (10) Lavender oil (11) Sodium alginate (1 2) Purified water (preparation method) (1) to (12) are mixed and dissolved according to a usual method to obtain a lotion. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 shows the activity of a raspberry extract and an elastase-based elastase. Fig. 2 is a graph showing the elastase inhibition and tongue properties of various pharmaceutical agents and mixtures thereof. 150170.doc .18-

Claims (1)

201206493 VII. Application for patent garden: It contains raspberry extract and hydroxy hydrazine 1 · An elastase inhibitor Amino acid as an active ingredient. 2. The elastase inhibitor of claim 1 is L-hydroxyproline. Wherein the above hydroxyproline acid further comprises yeast extract 3. As requested! Or 2 elastase inhibitors. 4. The elastase-inhibiting ❹ y 丨 preparation according to claim 3, wherein the yeast extract is cultured in a camping medium containing glycosaminoglycan I <® and irradiated with ultraviolet light, hydrogen peroxide or It is prepared by the yeast treated by the two temples. A skin external preparation comprising an elastase inhibitor as claimed in any one of claims 1 to 4 / y = 5 • 3 π items 1 to 4. 6. The skin external preparation of claim 5, which is a cosmetic. 〇 150170.doc